Microneedle devices and methods of manufacture and use thereof

ABSTRACT

Microneedle devices are provided for transport of therapeutic and biological molecules across tissue barriers and for use as microflameholders. In a preferred embodiment for transport across tissue, the microneedles are formed of a biodegradable polymer. Methods of making these devices, which can include hollow and/or porous microneedles, are also provided. A preferred method for making a microneedle includes forming a micromold having sidewalls which define the outer surface of the microneedle, electroplating the sidewalls to form the hollow microneedle, and then removing the micromold from the microneedle. In a preferred method of use, the microneedle device is used to deliver fluid material into or across a biological barrier from one or more chambers in fluid connection with at least one of the microneedles. The device preferably further includes a means for controlling the flow of material through the microneedles. Representative examples of these means include the use of permeable membranes, fracturable impermeable membranes, valves, and pumps.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a continuation-in-part of U.S. Ser. No. 09/095,221, filed Jun.10, 1998 now U.S. Pat. No. 6,193,700.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

The government has certain rights in this invention by virtue of GrantNumber BES-9624832 awarded by the U.S. National Science Foundation toMark R. Prausnitz, and by virtue of Contract Number DABT63-95-C-0114awarded by the U.S. Defense Advanced Research Projects Agency (DARPA).

BACKGROUND OF THE INVENTION

This invention is generally in the field of devices for the transport oftherapeutic or biological molecules across tissue barriers, such as fordrug delivery.

Numerous drugs and therapeutic agents have been developed in the battleagainst disease and illness. However, a frequent limitation of thesedrugs is their delivery: how to transport drugs across biologicalbarriers in the body (e.g., the skin, the oral mucosa, the blood-brainbarrier), which normally do not transport drugs at rates that aretherapeutically useful or optimal.

Drugs are commonly administered orally as pills or capsules. However,many drugs cannot be effectively delivered in this manner, due todegradation in the gastrointestinal tract and/or elimination by theliver. Moreover, some drugs cannot effectively diffuse across theintestinal mucosa. Patient compliance may also be a problem, forexample, in therapies requiring that pills be taken at particularintervals over a prolonged time.

Another common technique for delivering drugs across a biologicalbarrier is the use of a needle, such as those used with standardsyringes or catheters, to transport drugs across (through) the skin.While effective for this purpose, needles generally cause pain; localdamage to the skin at the site of insertion; bleeding, which increasesthe risk of disease transmission; and a wound sufficiently large to be asite of infection. The withdrawal of bodily fluids, such as fordiagnostic purposes, using a conventional needle has these samedisadvantages. Needle techniques also generally require administrationby one trained in its use. The needle technique also is undesirable forlong term, controlled continuous drug delivery.

Similarly, current methods of sampling biological fluids are invasiveand suffer from the same disadvantages. For example, needles are notpreferred for frequent routine use, such as sampling of a diabetic'sblood glucose or delivery of insulin, due to the vascular damage causedby repeated punctures. No alternative methodologies are currently inuse. Proposed alternatives to the needle require the use of lasers orheat to create a hole in the skin, which is inconvenient, expensive, orundesirable for repeated use.

An alternative delivery technique is the transdermal patch, whichusually relies on diffusion of the drug across the skin. However, thismethod is not useful for many drugs, due to the poor permeability (i.e.effective barrier properties) of the skin. The rate of diffusion dependsin part on the size and hydrophilicity of the drug molecules and theconcentration gradient across the stratum corneum. Few drugs have thenecessary physiochemical properties to be effectively delivered throughthe skin by passive diffusion. Iontophoresis, electroporation,ultrasound, and heat (so-called active systems) have been used in anattempt to improve the rate of delivery. While providing varying degreesof enhancement, these techniques are not suitable for all types ofdrugs, failing to provide the desired level of delivery. In some cases,they are also painful and inconvenient or impractical for continuouscontrolled drug delivery over a period of hours or days. Attempts havebeen made to design alternative devices for active transfer of drugs, oranalyte to be measured, through the skin.

For example, U.S. Pat. No. 5,879,326 to Godshall et al. and PCT WO96/37256 by Silicon Microdevices, Inc. disclose a transdermal drugdelivery apparatus that includes a cutter portion having a plurality ofmicroprotrusions, which have straight sidewalls, extending from asubstrate that is in communication with a drug reservoir. In operation,the microprotrusions penetrate the skin until limited by a stop regionof the substrate and then are moved parallel to the skin to createincisions. Because the microprotrusions are dragged across the skin, thedevice creates a wound sufficiently large to be a site of infection.Channels in the substrate adjacent to the microprotrusions allow drugfrom the reservoir to flow to the skin near the area disrupted by themicroprotrusions. Merely creating a wound, rather than using a needlewhich conveys drug through an enclosed channel into the site ofadministration, also creates more variability in dosage.

U.S. Pat. No. 5,250,023 to Lee et al. discloses a transdermal drugdelivery device, which includes a plurality of skin needles having adiameter in the range of 50 to 400 μm. The skin needles are supported ina water-swellable polymer substrate through which a drug solutionpermeates to contact the surface of the skin. An electric current isapplied to the device to open the pathways created by the skin needles,following their withdrawal from the skin upon swelling of the polymersubstrate.

PCT WO 93/17754 by Gross et al. discloses another transdermal drugdelivery device that includes a housing having a liquid drug reservoirand a plurality of tubular elements for transporting liquid drug intothe skin. The tubular elements may be in the form of hollow needleshaving inner diameters of less than 1 mm and an outer diameter of 1.0mm.

While each of these devices has potential use, there remains a need forbetter drug delivery devices, which make smaller incisions, deliver drugwith greater efficiency (greater drug delivery per quantity applied) andless variability of drug administration, and/or are easier to use.

It is therefore an object of the present invention to provide amicroneedle device for relatively painless, controlled, safe, convenienttransdermal delivery of a variety of drugs.

It is another object of the present invention to provide a microneedledevice for controlled sampling of biological fluids in aminimally-invasive, painless, and convenient manner.

It is still another object of the present invention to provide a hollowmicroneedle array for use in delivery or sensing of drugs or biologicalfluids or molecules.

SUMMARY OF THE INVENTION

Microneedle devices for transport of molecules, including drugs andbiological molecules, across tissue, and methods for manufacturing thedevices, are provided. The microneedle devices permit drug delivery orremoval of body fluids at clinically relevant rates across skin or othertissue barriers, with minimal or no damage, pain, or irritation to thetissue. Microneedles can be formed of biodegradable or non-biodegradablepolymeric materials or metals. In a preferred embodiment, themicroneedles are formed of a biodegradable polymer. In another preferredembodiment, the device includes a means for temporarily securing themicroneedle device to the biological barrier to facilitate transport.

Methods are provided for making porous or hollow microneedles. Apreferred method for making a microneedle includes forming a micromoldhaving sidewalls which define the outer surface of the microneedle. Themicromold can be formed, for example, by photolithographically definingone or more holes in a substrate, or by laser based cutting (eitherserially or by using lithographic projection), or by using amold-insert. In a preferred embodiment, the method includeselectroplating the sidewalls to form the hollow microneedle, and thenremoving the micromold from the microneedle.

The microneedle device is useful for delivery of fluid material into oracross a biological barrier wherein the fluid material is delivered fromone or more chambers in fluid connection with at least one of themicroneedles. The device preferably further includes a means forcontrolling the flow of material through the microneedles.Representative examples of these means include the use of permeablemembranes, fracturable impermeable membranes, valves, and pumps, andelectrical means.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1a is a side elevational view of a preferred embodiment of themicroneedle device inserted into human skin.

FIG. 1b is a diagram of one embodiment of microneedles.

FIGS. 2a-e are side cross-sectional views of a method for makingmicroneedles.

FIGS. 3a-g are side cross-sectional views of a method for making ahollow microneedle.

FIGS. 4a through 4 d are side cross-sectional views illustrating apreferred method for making hollow microneedles.

FIGS. 5a through 5 d are side cross-sectional views illustrating apreferred method for making hollow silicon microtubes.

FIGS. 6a through 6 e are side cross-sectional views illustrating apreferred method for making hollow metal microtubes.

FIGS. 7a through 7 d are side cross-sectional views illustrating apreferred method for making tapered metal microneedles.

FIGS. 8a through 8 d are side cross-sectional views illustrating amethod for making tapered microneedles using laser-formed molds.

FIGS. 9a through 9 f are side cross-sectional views illustrating asecond method for making tapered microneedles using laser-formed molds.

DETAILED DESCRIPTION OF THE INVENTION

1. Biological Barriers

The devices disclosed herein are useful in transport of material into oracross biological barriers including the skin (or parts thereof); theblood-brain barrier; mucosal tissue (e.g., oral, nasal, ocular, vaginal,urethral, gastrointestinal, respiratory); blood vessels; lymphaticvessels; or cell membranes (e.g., for the introduction of material intothe interior of a cell or cells). The biological barriers can be inhumans or other types of animals, as well as in plants, insects, orother organisms, including bacteria, yeast, fungi, and embryos.

The microneedle devices can be applied to tissue internally with the aidof a catheter or laparoscope. For certain applications, such as for drugdelivery to an internal tissue, the devices can be surgically implanted.

The microneedle device disclosed herein is typically applied to skin.The stratum corneum is the outer layer, generally between 10 and 50cells, or between 10 and 20 μm thick. Unlike other tissue in the body,the stratum corneum contains “cells” (called keratinocytes) filled withbundles of cross-linked keratin and keratohyalin surrounded by anextracellular matrix of lipids. It is this structure that is believed togive skin its barrier properties, which prevents therapeutic transdermaladministration of many drugs. Below the stratum corneum is the viableepidermis, which is between 50 and 100 μm thick. The viable epidermiscontains no blood vessels, and it exchanges metabolites by diffusion toand from the dermis. Beneath the viable epidermis is the dermis, whichis between 1 and 3 mm thick and contains blood vessels, lymphatics, andnerves.

2. The Microneedle Device

The microneedle devices disclosed herein include a substrate; one ormore microneedles; and, optionally, a reservoir for delivery of drugs orcollection of analyte, as well as pump(s), sensor(s), and/ormicroprocessor(s) to control the interaction of the foregoing.

a. Substrate

The substrate of the device can be constructed from a variety ofmaterials, including metals, ceramics, semiconductors, organics,polymers, and composites. The substrate includes the base to which themicroneedles are attached or integrally formed. A reservoir may also beattached to the substrate.

b. Microneedle

The microneedles of the device can be constructed from a variety ofmaterials, including metals, ceramics, semiconductors, organics,polymers, and composites. Preferred materials of construction includepharmaceutical grade stainless steel, gold, titanium, nickel, iron,gold, tin, chromium, copper, alloys of these or other metals, silicon,silicon dioxide, and polymers. Representative biodegradable polymersinclude polymers of hydroxy acids such as lactic acid and glycolic acidpolylactide, polyglycolide, polylactide-co-glycolide, and copolymerswith PEG, polyanhydrides, poly(ortho)esters, polyurethanes, poly(butyricacid), poly(valeric acid), and poly(lactide-co-caprolactone).Representative non-biodegradable polymers include polycarbonate,polymethacrylic acid, ethylenevinyl acetate, polytetrafluorethylene(TEFLON™), and polyesters.

Generally, the microneedles should have the mechanical strength toremain intact for delivery of drugs, or serve as a conduit for thecollection of biological fluid, while being inserted into the skin,while remaining in place for up to a number of days, and while beingremoved. In embodiments where the microneedles are formed ofbiodegradable polymers, however, this mechanical requirement is lessstringent, since the microneedles or tips thereof can break off, forexample in the skin, and will biodegrade. Nonetheless, even abiodegradable microneedle still needs to remain intact at least longenough for the microneedle to serve its intended purpose (e.g, itsconduit function). Therefore, biodegradable microneedles can provide anincreased level of safety, as compared to nonbiodegradable ones. Themicroneedles should be sterilizable using standard methods.

The microneedles can be formed of a porous solid, with or without asealed coating or exterior portion, or hollow. As used herein, the term“porous” means having pores or voids throughout at least a portion ofthe microneedle structure, sufficiently large and sufficientlyinterconnected to permit passage of fluid and/or solid materials throughthe microneedle. As used herein, the term “hollow” means having one ormore substantially annular bores or channels through the interior of themicroneedle structure, having a diameter sufficiently large to permitpassage of fluid and/or solid materials through the microneedle. Theannular bores may extend throughout all or a portion of the needle inthe direction of the tip to the base, extending parallel to thedirection of the needle or branching or exiting at a side of the needle,as appropriate. A solid or porous microneedle can be hollow. One ofskill in the art can select the appropriate porosity and/or borefeatures required for specific applications. For example, one can adjustthe pore size or bore diameter to permit passage of the particularmaterial to be transported through the microneedle device.

The microneedles can have straight or tapered shafts. A hollowmicroneedle that has a substantially uniform diameter, which needle doesnot taper to a point, is referred to herein as a “microtube.” As usedherein, the term “microneedle” includes both microtubes and taperedneedles unless otherwise indicated. In a preferred embodiment, thediameter of the microneedle is greatest at the base end of themicroneedle and tapers to a point at the end distal the base. Themicroneedle can also be fabricated to have a shaft that includes both astraight (untapered) portion and a tapered portion.

The microneedles can be formed with shafts that have a circularcross-section in the perpendicular, or the cross-section can benon-circular. For example, the cross-section of the microneedle can bepolygonal (e.g. star-shaped, square, triangular), oblong, or anothershape. The shaft can have one or more bores. The cross-sectionaldimensions typically are between about 10 nm and 1 mm, preferablybetween 1 micron and 200 microns, and more preferably between 10 and 100μm. The outer diameter is typically between about 10 μm and about 100μm, and the inner diameter is typically between about 3 μm and about 80μm.

The length of the microneedles typically is between about 1 μm and 1 mm,preferably between 10 microns and 500 microns, and more preferablybetween 30 and 200 μm. The length is selected for the particularapplication, accounting for both an inserted and uninserted portion. Anarray of microneedles can include a mixture of microneedles having, forexample, various lengths, outer diameters, inner diameters,cross-sectional shapes, and spacings between the microneedles.

The microneedles can be oriented perpendicular or at an angle to thesubstrate. Preferably, the microneedles are oriented perpendicular tothe substrate so that a larger density of microneedles per unit area ofsubstrate can be provided. An array of microneedles can include amixture of microneedle orientations, heights, or other parameters.

In a preferred embodiment of the device, the substrate and/ormicroneedles, as well as other components, are formed from flexiblematerials to allow the device to fit the contours of the biologicalbarrier, such as the skin, vessel walls, or the eye, to which the deviceis applied. A flexible device will facilitate more consistentpenetration during use, since penetration can be limited by deviationsin the attachment surface. For example, the surface of human skin is notflat due to dermatoglyphics (i.e. tiny wrinkles) and hair.

c. Reservoir

The microneedle device may include a reservoir in communication with themicroneedles. The reservoir can be attached to the substrate by anysuitable means. In a preferred embodiment, the reservoir is attached tothe back of the substrate (opposite the microneedles) around theperiphery, using an adhesive agent (e.g., glue). A gasket may also beused to facilitate formation of a fluid-tight seal.

In a preferred embodiment, the reservoir contains drug, for deliverythrough the microneedles. The reservoir may be a hollow vessel, a porousmatrix, or a solid form including drug which is transported therefrom.The reservoir can be formed from a variety of materials that arecompatible with the drug or biological fluid contained therein.Preferred materials include natural and synthetic polymers, metals,ceramics, semiconductors, organics, and composites.

The microneedle device can include one or a plurality of chambers forstoring materials to be delivered. In the embodiment having multiplechambers, each can be in fluid connection with all or a portion of themicroneedles of the device array. In one embodiment, at least twochambers are used to separately contain drug (e.g., a lyophilized drug,such as a vaccine) and an administration vehicle (e.g., saline) in orderto prevent or minimize degradation during storage. Immediately beforeuse, the contents of the chambers are mixed. Mixing can be triggered byany means, including, for example, mechanical disruption (i.e.puncturing or breaking), changing the porosity, or electrochemicaldegradation of the walls or membranes separating the chambers. Inanother embodiment, a single device is used to deliver different drugs,which are stored separately in different chambers. In this embodiment,the rate of delivery of each drug can be independently controlled.

In a preferred embodiment, the reservoir should be in direct contactwith the microneedles and have holes through which drug could exit thereservoir and flow into the interior of hollow or porous microneedles.In another preferred embodiment, the reservoir has holes which permitthe drug to transport out of the reservoir and onto the skin surface.From there, drug is transported into the skin, either through hollow orporous microneedles, along the sides of solid microneedles, or throughpathways created by microneedles in the skin.

d. Transport Control Components

The microneedle device also must be capable of transporting materialacross the barrier at a useful rate. For example, the microneedle devicemust be capable of delivering drug across the skin at a rate sufficientto be therapeutically useful. The device may include a housing withmicroelectronics and other micromachined structures to control the rateof delivery either according to a preprogrammed schedule or throughactive interface with the patient, a healthcare professional, or abiosensor. The rate can be controlled by manipulating a variety offactors, including the characteristics of the drug formulation to bedelivered (e.g., its viscosity, electric charge, and chemicalcomposition); the dimensions of each microneedle (e.g., its outerdiameter and the area of porous or hollow openings); the number ofmicroneedles in the device; the application of a driving force (e.g., aconcentration gradient, a voltage gradient, a pressure gradient); andthe use of a valve.

The rate also can be controlled by interposing between the drug in thereservoir and the opening(s) at the base end of the microneedlepolymeric or other materials selected for their diffusioncharacteristics. For example, the material composition and layerthickness can be manipulated using methods known in the art to vary therate of diffusion of the drug of interest through the material, therebycontrolling the rate at which the drug flows from the reservoir throughthe microneedle and into the tissue.

Transportation of molecules through the microneedles can be controlledor monitored using, for example, various combinations of valves, pumps,sensors, actuators, and microprocessors. These components can beproduced using standard manufacturing or microfabrication techniques.Actuators that may be useful with the microneedle devices disclosedherein include micropumps, microvalves, and positioners. In a preferredembodiment, a microprocessor is programmed to control a pump or valve,thereby controlling the rate of delivery.

Flow of molecules through the microneedles can occur based on diffusion,capillary action, or can be induced using conventional mechanical pumpsor nonmechanical driving forces, such as electroosmosis orelectrophoresis, or convection. For example, in electroosmosis,electrodes are positioned on the biological barrier surface, one or moremicroneedles, and/or the substrate adjacent the needles, to create aconvective flow which carries oppositely charged ionic species and/orneutral molecules toward or into the biological barrier. In a preferredembodiment, the microneedle device is used in combination with anothermechanism that enhances the permeability of the biological barrier, forexample by increasing cell uptake or membrane disruption, using electricfields, ultrasound, chemical enhancers, viruses, pH, heat and/or light.

Passage of the microneedles, or drug to be transported via themicroneedles, can be manipulated by shaping the microneedle surface, orby selection of the material forming th microneedle surface (which couldbe a coating rather than the microneedle per se). For example, one ormore grooves on the outside surface of the microneedles can be used todirect the passage of drug, particularly in a liquid state.Alternatively, the physical surface properties of the microneedle couldbe manipulated to either promote or inhibit transport of material alongthe microneedle surface, such as by controlling hydrophilicity orhydrophobicity.

The flow of molecules can be regulated using a wide range of valves orgates. These valves can be the type that are selectively and repeatedlyopened and closed, or they can be single-use types. For example, in adisposable, single-use drug delivery device, a fracturable barrier orone-way gate may be installed in the device between the reservoir andthe opening of the microneedles. When ready to use, the barrier can bebroken or gate opened to permit flow through the microneedles. Othervalves or gates used in the microneedle devices can be activatedthermally, electrochemically, mechanically, or magnetically toselectively initiate, modulate, or stop the flow of molecules throughthe needles. In a preferred embodiment, flow is controlled by using arate-limiting membrane as a “valve.”

The microneedle devices can further include a flowmeter or other meansto monitor flow through the microneedles and to coordinate use of thepumps and valves.

e. Sensors

Useful sensors may include sensors of pressure, temperature, chemicals,and/or electromagnetic fields. Biosensors can be located on themicroneedle surface, inside a hollow or porous microneedle, or inside adevice in communication with the body tissue via the microneedle (solid,hollow, or porous). These microneedle biosensors can include fourclasses of principal transducers: potentiometric, amperometric, optical,and physiochemical. An amperometric sensor monitors currents generatedwhen electrons are exchanged between a biological system and anelectrode. Blood glucose sensors frequently are of this type.

The microneedle may function as a conduit for fluids, solutes, electriccharge, light, or other materials. In one embodiment, hollowmicroneedles can be filled with a substance, such as a gel, that has asensing functionality associated with it. In an application for sensingbased on binding to a substrate or reaction mediated by an enzyme, thesubstrate or enzyme can be immobilized in the needle interior, whichwould be especially useful in a porous needle to create an integralneedle/sensor.

Wave guides can be incorporated into the microneedle device to directlight to a specific location, or for dection, for example, using meanssuch as a pH dye for color evaluation. Similarly, heat, electricity,light or other energy forms may be precisely transmitted to directlystimulate, damage, or heal a specific tissue or intermediary (e.g.,tattoo remove for dark skinned persons), or diagnostic purposes, such asmeasurement of blood glucose based on IR spectra or by chromatographicmeans, measuring a color change in the presence of immobilized glucoseoxidase in combination with an appropriate substrate.

f. Attachment Features

A collar or flange also can be provided with the device, for example,around the periphery of the substrate or the base. It preferably isattached to the device, but alternatively can be formed as integral partof the substrate, for example by forming microneedles only near thecenter of an “oversized” substrate. The collar can also emanate fromother parts of the device. The collar can provide an interface to attachthe microneedle array to the rest of the device, and can facilitatehandling of the smaller devices.

In a preferred embodiment, the microneedle device includes an adhesiveto temporarily secure the device to the surface of the biologicalbarrier. The adhesive can be essentially anywhere on the device tofacilitate contact with the biological barrier. For example, theadhesive can be on the surface of the collar (same side asmicroneedles), on the surface of the substrate between the microneedles(near the base of the microneedles), or a combination thereof.

g. Transdermal Microneedle Device

FIG. 1a is a side elevational view of a schematic of a preferredembodiment of the microneedle device inserted into skin. The device 10includes an upper portion or substrate 11 from which a plurality ofmicroneedles 12 protrude. The height of the upper portion 11 is betweenabout 1 μm and 1 cm, and the width of the upper portion is between about1 mm and 10 cm. The upper portion 11 of the device can be solid orhollow, and may include multiple compartments. In a preferred embodimentfor drug delivery, the upper portion 11 contains one or more drugs to bedelivered. It is also preferred that the upper portion include one ormore sensors and/or an apparatus (e.g., pump or electrode) to drive(provide/direct the force) transport of the drug or other molecules.

The height (or length) of the microneedles 12 generally is between about1 μm and 1 mm. The diameter and length both affect pain as well asfunctional properties of the needles. In transdermal applications, the“insertion depth” of the microneedles 12 is preferably less than about100 μm, more preferably about 30 μm, so that insertion of themicroneedles 12 into the skin through the stratum corneum 14 does notpenetrate past the epidermis 16 into the dermis 18 (as described below),thereby avoiding contacting nerves and reducing the potential forcausing pain. In such applications, the actual length of themicroneedles may be longer, since the portion of the microneedles distalthe tip may not be inserted into the skin; the uninserted length dependson the particular device design and configuration. The actual (overall)height or length of microneedles 12 should be equal to the insertiondepth plus the uninserted length.

The diameter of each microneedle 12 generally is between about 10 nm and1 mm, and preferably leaves a residual hole (following microneedleinsertion and withdrawal) of less than about 1 μm, to avoid making ahole which would allow bacteria to enter the penetration wound. Theactual microneedle diameter should be larger than 1 μm, since the holelikely will contract following withdrawal of the microneedle. Thediameter of microneedle 12 more preferably is between about 1 μm and 100μm. Larger diameter and longer microneedles are acceptable, so long asthe microneedle can penetrate the biological barrier to the desireddepth and the hole remaining in the skin or other tissue followingwithdrawal of the microneedle is sufficiently small, preferably smallenough to exclude bacterial entry. The microneedles 12 can be solid orporous, and can include one or more bores connected to upper portion 11.

3. Methods of Making Microneedle Devices

The microneedle devices are made by microfabrication processes, bycreating small mechanical structures in silicon, metal, polymer, andother materials. These microfabrication processes are based onwell-established methods used to make integrated circuits, electronicpackages and other microelectronic devices, augmented by additionalmethods used in the field of micromachining. The microneedle devices canhave dimensions as small as a few nanometers and can be mass-produced atlow per-unit costs.

a. Microfabrication Processes

Microfabrication processes that may be used in making the microneedlesdisclosed herein include lithography; etching techniques, such as wetchemical, dry, and photoresist removal; thermal oxidation of silicon;electroplating and electroless plating; diffusion processes, such asboron, phosphorus, arsenic, and antimony diffusion; ion implantation;film deposition, such as evaporation (filament, electron beam, flash,and shadowing and step coverage), sputtering, chemical vapor deposition(CVD), epitaxy (vapor phase, liquid phase, and molecular beam),electroplating, screen printing, lamination, stereolithography, lasermachining, and laser ablation (including projection ablation). Seegenerally Jaeger, Introduction to Microelectronic Fabrication(Addison-Wesley Publishing Co., Reading Mass. 1988); Runyan, et al.,Semiconductor Integrated Circuit Processing Technology (Addison-WesleyPublishing Co., Reading Mass. 1990); Proceedings of the IEEE MicroElectro Mechanical Systems Conference 1987-1998; Rai-Choudhury, ed.,Handbook of Microlithography, Micromachining & Microfabrication (SPIEOptical Engineering Press, Bellingham, Wash. 1997).

The following methods are preferred for making microneedles.

i. Electrochemical Etching of Silicon

In this method, electrochemical etching of solid silicon to poroussilicon is used to create extremely fine (on the order of 0.01 μm)silicon networks which can be used as piercing structures. This methoduses electrolytic anodization of silicon in aqueous hydrofluoric acid,potentially in combination with light, to etch channels into thesilicon. By varying the doping concentration of the silicon wafer to beetched, the electrolytic potential during etching, the incident lightintensity, and the electrolyte concentration, control over the ultimatepore structure can be achieved. The material not etched (i.e. thesilicon remaining) forms the microneedles. This method has been used toproduce irregular needle-type structures measuring tens of nanometers inwidth.

ii. Plasma Etching

This process uses deep plasma etching of silicon to create microneedleswith diameters on the order of 0.1 μm or larger. Needles are patterneddirectly using photolithography, rather than indirectly by controllingthe voltage (as in electrochemical etching), thus providing greatercontrol over the final microneedle geometry.

In this process, an appropriate masking material (e.g., metal) isdeposited onto a silicon wafer substrate and patterned into dots havingthe diameter of the desired microneedles. The wafer is then subjected toa carefully controlled plasma based on fluorine/oxygen chemistries toetch very deep, high aspect ratio trenches into the silicon. See, e.g.,Jansen, et al., “The Black Silicon Method IV: The Fabrication ofThree-Dimensional Structures in Silicon with High Aspect Ratios forScanning Probe Microscopy and Other Applications,” IEEE ProceedingsofMicro Electro Mechanical Systems Conference, pp. 88-93 (1995). Thoseregions protected by the metal mask remain and form the needles. Thismethod is further described in Example 1 below. FIG. 1b provides adiagram of microneedles fabricated by this method.

iii. Electroplating

In this process, a metal layer is first evaporated onto a planarsubstrate. A layer of photoresist is then deposited onto the metal toform a patterned mold which leaves an exposed-metal region in the shapeof needles. By electroplating onto the exposed regions of the metal seedlayer, the mold bounded by photoresist can be filled with electroplatedmaterial. Finally, the substrate and photoresist mold are removed,leaving the finished microneedle array. The microneedles produced bythis process generally have diameters on the order of 1 μm or larger.See, e.g., Frazier, et al., “Two dimensional metallic microelectrodearrays for extracellular stimulation and recording of neurons”, IEEEProceedings of the Micro Electro Mechanical Systems Conference, pp.195-200 (1993).

iv. Other Processes

Another method for forming microneedles made of silicon or othermaterials is to use microfabrication techniques such asphotolithography, plasma etching, or laser ablation to make a mold form(A), transferring that mold form to other materials using standard moldtransfer techniques, such as embossing or injection molding (B), andreproducing the shape of the original mold form (A) using thenewly-created mold (B) to yield the final microneedles (C).Alternatively, the creation of the mold form (A) could be skipped andthe mold (B) could be microfabricated directly, which could then be usedto create the final microneedles (C).

Another method of forming solid silicon microneedles is by usingepitaxial growth on silicon substrates, as is utilized by ContainerlessResearch, Inc. (Evanston, Ill., USA) for its products.

b. Hollow or Porous Microneedles

In a preferred embodiment, microneedles are made with pores or otherpathways through which material may be transported. The followingdescriptions outline representative methods for fabricating eitherporous or hollow microneedles.

i. Porous Microneedles

Rather than having a single, well-defined hole down the length of theneedle, porous needles are filled with a network of channels or poreswhich allow conduction of fluid or energy through the needle shaft. Ithas been shown that by appropriate electrochemical oxidation of silicon,pore arrays with high aspect ratios and a range of different pore sizeregimes can be formed; these pore regimes are defined as (1) microporousregime with average pore dimensions less than 2 nm, (2) mesoporousregime with average pore sizes of between 2 nm and 50 nm, and (3)macroporous regime with pores greater than 50 nm. The mesoporous andmacroporous regimes are expected to be most useful for drug delivery.Two approaches to porous needles are generally available, either (a) thesilicon wafer is first made porous and then etched as described above toform needles or (b) solid microneedles are etched and then renderedporous, for example, by means of electrochemical oxidation, such as byanodization of a silicon substrate in a hydrofluoric acid electrolyte.The size distribution of the etched porous structure is highly dependenton several variables, including doping kind and illumination conditions,as detailed in Lehmann, “Porous Silicon—A New Material for MEMS”, IEEEProceedings of the Micro Electro Mechanical Systems Conference, pp. 1-6(1996). Porous polymer or metallic microneedles can be formed, forexample, by micromolding a polymer containing a volatilizable orleachable material, such as a volatile salt, dispersed in the polymer ormetal, and then volatilizing or leaching the dispersed material, leavinga porous polymer matrix in the shape of the microneedle.

ii. Hollow Needles

Three-dimensional arrays of hollow microneedles can be fabricated, forexample, using combinations of dry etching processes (Laermer, et al.,“Bosch Deep Silicon Etching: Improving Uniformity and Etch Rate forAdvanced MEMS Applications,” Micro Electro Mechanical Systems, Orlando,Fla., USA, (Jan. 17-21, 1999); Despont et al., “High-Aspect-Ratio,Ultrathick, Negative-Tone Near-UV Photoresist for MEMS”, Proc. of IEEE10^(th) Annual International Workshop on MEMS, Nagoya, Japan, pp.518-522 (Jan. 26-30, 1997)); micromold creation inlithographically-defined and/or laser ablated polymers and selectivesidewall electroplating; or direct micromolding techniques using epoxymold transfers.

One or more distinct and continuous pathways are created through theinterior of microneedles. In a preferred embodiment, the microneedle hasa single annular pathway along the center axis of the microneedle. Thispathway can be achieved by initially chemically or physically etchingthe holes in the material and then etching away microneedles around thehole. Alternatively, the microneedles and their holes can be madesimultaneously or holes can be etched into existing microneedles. Asanother option, a microneedle form or mold can be made, then coated, andthen etched away, leaving only the outer coating to form a hollowmicroneedle. Coatings can be formed either by deposition of a film or byoxidation of the silicon microneedles to a specific thickness, followedby removal of the interior silicon. Also, holes from the backside of thewafer to the underside of the hollow needles can be created using afront-to-backside infrared alignment followed by etching from thebackside of the wafer.

a. Silicon Microneedles

One method for hollow needle fabrication is to replace the solid maskused in the formation of solid needles by a mask that includes a solidshape with one or more interior regions of the solid shape removed. Oneexample is a “donut-shaped” mask. Using this type of mask, interiorregions of the needle are etched simultaneously with their side walls.Due to lateral etching of the inner side walls of the needle, this maynot produce sufficiently sharp walls. In that case, two plasma etchesmay be used, one to form the outer walls of the microneedle (i.e., the‘standard’ etch), and one to form the inner hollow core (which is anextremely anisotropic etch, such as in inductively-coupled-plasma “ICP”etch). For example, the ICP etch can be used to form the interior regionof the needle followed by a second photolithography step and a standardetch to form the outer walls of the microneedle. FIG. 2a represents asilicon wafer 82 with a patterned photoresist layer 84 on top of thewafer 82. The wafer 82 is anisotropically etched (FIG. 2b) to form acavity 86 through its entire thickness (FIG. 2c). The wafer 82 is thencoated with a chromium layer 88 followed by a second photoresist layer90 patterned so as to cover the cavity 86 and form a circular mask forsubsequent etching (FIG. 2d). The wafer 82 is then etched by a standardetch to form the outer tapered walls 92 of the microneedle (FIG. 2e).

Alternatively, this structure can be achieved by substituting thechromium mask used for the solid microneedles described in Example I bya silicon nitride layer 94 on the silicon substrate 95 covered withchromium 96, deposited as shown in FIG. 3a and patterned as shown inFIG. 3b. Solid microneedles are then etched as described in Example 1 asshown FIG. 3c, the chromium 96 is stripped (FIG. 3d), and the silicon 95is oxidized to form a thin layer of silicon dioxide 97 on all exposedsilicon surfaces (FIG. 3e). The silicon nitride layer 94 preventsoxidation at the needle tip. The silicon nitride 94 is then stripped(FIG. 3f), leaving exposed silicon at the tip of the needle andoxide-covered silicon 97 everywhere else. The needle is then exposed toan ICP plasma which selectively etches the inner sidewalls of thesilicon 95 in a highly anisotropic manner to form the interior hole ofthe needle (FIG. 3g).

Another-method uses the solid silicon needles described previously as‘forms’ around which the actual needle structures are deposited. Afterdeposition, the forms are etched away, yielding the hollow structures.Silica needles or metal needles can be formed using different methods.Silica needles can be formed by creating needle structures similar tothe ICP needles described above prior to the oxidation described above.The wafers are then oxidized to a controlled thickness, forming a layeron the shaft of the needle form which will eventually become the hollowmicroneedle. The silicon nitride is then stripped and the silicon coreselectively etched away (e.g., in a wet alkaline solution) to form ahollow silica microneedle.

In a preferred embodiment, an array of hollow silicon microtubes is madeusing deep reactive ion etching combined with a modified black siliconprocess in a conventional reactive ion etcher, as described in Example 3below. First, arrays of circular holes are patterned through photoresistinto SiO₂, such as on a silicon wafer. Then the silicon can be etchedusing deep reactive ion etching (DRIE) in an inductively coupled plasma(ICP) reactor to etch deep vertical holes. The photoresist was thenremoved. Next, a second photolithography step patterns the remainingSiO₂ layer into circles concentric to the holes, leaving ring shapedoxide masks surrounding the holes. The photoresist is then removed andthe silicon wafer again deep silicon etched, such that the holes areetched completely through the wafer (inside the SiO₂ ring) andsimultaneously the silicon is etched around the SiO₂ ring leaving acylinder.

This latter process can be varied to produce hollow, taperedmicroneedles. After an array of holes is fabricated as described above,the photoresist and SiO₂ layers are replaced with conformal DC sputteredchromium rings. The second ICP etch is replaced with a SF₆/O₂ plasmaetch in a reactive ion etcher (RIE), which results in positively slopingouter sidewalls. Henry, et al., “Micromachined Needles for theTransdermal Delivery of Drugs,” Micro Electro Mechanical Systems,Heidelberg, Germany, pp. 494-498 (Jan. 26-29, 1998).

b. Metal Microneedles

Metal needles can be formed by physical vapor deposition of appropriatemetal layers on solid needle forms, which can be made of silicon usingthe techniques described above, or which can be formed using otherstandard mold techniques such as embossing or injection molding. Themetals are selectively removed from the tips of the needles usingelectropolishing techniques, in which an applied anodic potential in anelectrolytic solution will cause dissolution of metals more rapidly atsharp points, due to concentration of electric field lines at the sharppoints. Once the underlying silicon needle forms have been exposed atthe tips, the silicon is selectively etched away to form hollow metallicneedle structures. This process could also be used to make hollowneedles made from other materials by depositing a material other thanmetal on the needle forms and following the procedure described above.

A preferred method of fabricating hollow metal microneedles utilizesmicromold plating techniques, for example which are described as followsand in Examples 4 and 5. In a method for making metal microtubes, whichdoes not require dry silicon etching, a photo-defined mold first isfirst produced, for example, by spin casting a thick layer, typically150 μm, of an epoxy (e.g., SU-8) onto a substrate that has been coatedwith a thin sacrificial layer, typically about 10 to 50 nm. Arrays ofcylindrical holes are then photolithographically defined through theepoxy layer, which typically is about 150 μm thick. (Despont, et al.,“High-Aspect-Ratio, Ultrathick, Negative-Tone Near-UV Photoresist forMEMS,” Proc. of IEEE 10^(th) Annual International Workshop on MEMS,Nagoya, Japan, pp. 518-522 (Jan. 26-30, 1997)). The diameter of thesecylindrical holes defines the outer diameter of the tubes. The uppersurface of the substrate, the sacrificial layer, is then partiallyremoved at the bottom of the cylindrical holes in the photoresist. Theexact method chosen depends on the choice of substrate. For example, theprocess has been successfully performed on silicon and glass substrates(in which the upper surface is etched using isotropic wet or dry etchingtechniques) and copper-clad printed wiring board substrates. In thelatter case, the copper laminate is selectively removed using wetetching. Then a seed layer, such as Ti/Cu/Ti (e.g., 30 nm/200 nm/30 nm),is conformally DC sputter-deposited onto the upper surface of the epoxymold and onto the sidewalls of the cylindrical holes. The seed layershould be electrically isolated from the substrate. Subsequently, one ormore electroplatable metals or alloys, such as Ni, NiFe, Au, Cu, or Tiare electroplated onto the seed layer. The surrounding epoxy is thenremoved, leaving microtubes which each have an interior annular holethat extends through the base metal supporting the tubes. The rate andduration of electroplating is controlled in order to define the wallthickness and inner diameter of the microtubes. In one embodiment, thismethod was used to produce microtubes having a height of between about150 and 250 μm, an outer diameter of between about 40 and 120 μm, and aninner diameter of between about 30 and 110 μm (i.e., having a wallthickness of 10 μm). In a typical array, the microtubes have a tubecenter-to-center spacing of about 150 μm, but can vary depending on thedesired needle density.

A variation of this method is preferred for forming taperedmicroneedles. As described above, photolithography yields holes in theepoxy which have vertical sidewalls, such that the resulting shafts ofthe microneedles are straight, not tapered. This vertical sidewalllimitation can be overcome by molding a preexisting 3D structure, i.e.,a mold-insert. The subsequent removal of the mold-insert leaves a moldwhich can be surface plated similarly to the holes produced byphotolithography described above.

Alternatively, non-vertical sidewalls can be produced directly in thepolymeric mold into which electroplating will take place. For example,conventional photoresists known in the art can be exposed and developedin such as way as to have the surface immediately adjacent to the maskbe wider than the other surface. Specialized greyscale photoresists incombination with greyscale masks can accomplish the same effect.Laser-ablated molds can also be made with tapered sidewalls, e.g., byoptical adjustment of the beam (in the case of serial hole fabrication)or of the reticle or mold during ablation (in the case of projectionablation).

To form hollow tapered microneedles, the mold-insert is an array ofsolid silicon microneedles, formed as described in Henry, et al.,“Micromachined Needles for the Transdermal Delivery of Drugs,” MicroElectro Mechanical Systems, Heidelberg, Germany, Jan. 26-29, pp. 494-498(1998). First, a layer of a material, such as an epoxy (e.g., SU-8), isspin cast onto the array of silicon microneedles to completely blanketthe entire array. The epoxy settles during pre-bake to create a planarsurface above the silicon needle tips; the material is then fullypre-baked, photolithographically cross-linked, and post-baked.

The upper surface of the epoxy is then etched away, for example with anO₂/CHF₃ plasma, until the needle tips are exposed, preferably leavingbetween about 1 and 5 μm of tip protruding from the epoxy. The siliconis then selectively removed, for example by using a SF₆ plasma or aHNO_(3/)HF solution. The remaining epoxy micromold is the negative ofthe microneedles and has a small diameter hole where the tip of themicroneedle formerly protruded.

After the removal of the silicon, a seed layer, such as Ti—Cu—Ti isconformally sputter-deposited onto the epoxy micromold. Following thesame process sequence described for hollow metal microtubes, one or moreelectroplatable metals or alloys, such as Ni, NiFe, Au, or Cu, areelectroplated onto the seed layer. Finally, the epoxy is removed, forexample by using an O₂/CHF₃ plasma, leaving an array of hollow metalmicroneedles. In a preferred embodiment, this method is used to producemicroneedles having a height of between about 150 and 250 μm, an outerdiameter of between about 40 and 120 μm, and an inner diameter ofbetween about 50 and 100 μm. In a typical array, the microtubes have atube center-to-center spacing of about 150 μm, but can vary depending onthe desired needle density. The microneedles are 150 μm in height with abase diameter of 80 μm, a tip diameter of 10 μm, and a needle to needlespacing of 150 μm.

c. Silicon Dioxide Microneedles

Hollow microneedles formed of silicon dioxide can be made by oxidizingthe surface of the silicon microneedle forms (as described above),rather than depositing a metal and then etching away the solid needleforms to leave the hollow silicon dioxide structures. This method isillustrated in FIGS. 4a-4 d. FIG. 4a shows an array 24 of needle forms26 with masks 28 on their tips. In FIG. 4b, the needle forms 26 havebeen coated with a layer 30 of metal, silicon dioxide or other material.FIG. 4c shows the coated needle forms 26 with the masks 28 removed.Finally, in FIG. 4d, the needle forms 26 have been etched away, leavinghollow needles 30 made of metal, silicon dioxide, or other materials.

In one embodiment, hollow, porous, or solid microneedles are providedwith longitudinal grooves or other modifications to the exterior surfaceof the microneedles. Grooves, for example, should be useful in directingthe flow of molecules along the outside of microneedles.

d. Polymer Microneedles

In a preferred method, polymeric microneedles are made usingmicrofabricated molds. For example, the epoxy molds can be made asdescribed above and injection molding techniques can be applied to formthe microneedles in the molds (Weber, et al., “Micromolding—a powerfultool for the large scale production of precise microstructures”, Proc.SPIE—International Soc. Optical Engineer. 2879, 156-167 (1996); Schift,et al., “Fabrication of replicated high precision insert elements formicro-optical bench arrangements” Proc. SPIE—International Soc. OpticalEngineer. 3513, 122-134 (1998). These micromolding techniques arepreferred over other techniques described herein, since they can providerelatively less expensive replication, i.e. lower cost of massproduction. In a preferred embodiment, the polymer is biodegradable.

4. Microneedle Device Applications

The device may be used for single or multiple uses for rapid transportacross a biological barrier or may be left in place for longer times(e.g., hours or days) for long-term transport of molecules. Depending onthe dimensions of the device, the application site, and the route inwhich the device is introduced into (or onto) the biological barrier,the device may be used to introduce or remove molecules at specificlocations.

As discussed above, FIG. 1 shows a side elevational view of a schematicof a preferred embodiment of the microneedle device 10 in a transdermalapplication. The device 10 is applied to the skin such that themicroneedles 12 penetrate through the stratum corneum and enter theviable epidermis so that the tip of the microneedle at least penetratesinto the viable epidermis. In a preferred embodiment, drug molecules ina reservoir within the upper portion 11 flow through or around themicroneedles and into the viable epidermis, where the drug moleculesthen diffuse into the dermis for local treatment or for transportthrough the body.

To control the transport of material out of or into the device throughthe microneedles, a variety of forces or mechanisms can be employed.These include pressure gradients, concentration gradients, electricity,ultrasound, receptor binding, heat, chemicals, and chemical reactions.Mechanical or other gates in conjunction with the forces and mechanismsdescribed above can be used to selectively control transport of thematerial.

In particular embodiments, the device should be “user-friendly.” Forexample, in some transdermal applications, affixing the device to theskin should be relatively simple, and not require special skills. Thisembodiment of a microneedle may include an array of microneedlesattached to a housing containing drug in an internal reservoir, whereinthe housing has a bioadhesive coating around the microneedles. Thepatient can remove a peel-away backing to expose an adhesive coating,and then press the device onto a clean part of the skin, leaving it toadminister drug over the course of, for example, several days.

a. Drug Delivery

Essentially any drug or other bioactive agents can be delivered usingthese devices. Drugs can be proteins, enzymes, polysaccharides,polynucleotide molecules, and synthetic organic and inorganic compounds.A preferred drug is insulin. Representative agents includeanti-infectives, hormones, growth regulators, drugs regulating cardiacaction or blood flow, and drugs for pain control. The drug can be forlocal treatment or for regional or systemic therapy. The following arerepresentative examples, and disorders they are used to treat:

Calcitonin, osteoporosis

Enoxaprin, anticoagulant

Etanercept, rheumatoid arthritis

Erythropoietin, anemia

Fentanyl, postoperative and chronic pain

Filgrastin, low white blood cells from chemotherapy

Heparin, anticoagulant

Insulin, human, diabetes

Interferon Beta I a, multiple sclerosis

Lidocaine, local anesthesia

Somatropin, growth hormone

Sumatriptan, migraine headaches

In this way, many drugs can be delivered at a variety of therapeuticrates. The rate can be controlled by varying a number of design factors,including the outer diameter of the microneedle, the number and size ofpores or channels in each microneedle, the number of microneedles in anarray, the magnitude and frequency of application of the force drivingthe drug through the microneedle and/or the holes created by themicroneedles. For example, devices designed to deliver drug at differentrates might have more microneedles for more rapid delivery and fewermicroneedles for less rapid delivery. As another example, a devicedesigned to deliver drug at a variable rate could vary the driving force(e.g., pressure gradient controlled by a pump) for transport accordingto a schedule which was pre-programmed or controlled by, for example,the user or his doctor. The devices can be affixed to the skin or othertissue to deliver drugs continuously or intermittently, for durationsranging from a few seconds to several hours or days.

One of skill in the art can measure the rate of drug delivery forparticular microneedle devices using in vitro and in vivo methods knownin the art. For example, to measure the rate of transdermal drugdelivery, human cadaver skin mounted on standard diffusion chambers canbe used to predict actual rates. See Hadgraft & Guy, eds., TransdermalDrug Delivery: Developmental Issues and Research Initiatives (MarcelDekker, New York 1989); Bronaugh & Maibach, Percutaneous Absorption,Mechanisms—Methodology—Drug Delivery (Marcel Dekker, New York 1989).After filling the compartment on the dermis side of the diffusionchamber with saline, a microneedle array is inserted into the stratumcorneum; a drug solution is placed in the reservoir of the microneedledevice; and samples of the saline solution are taken over time andassayed to determine the rates of drug transport.

In an alternate embodiment, biodegradable or non-biodegradablemicroneedles can be used as the entire drug delivery device, wherebiodegradable microneedles are a preferred embodiment. For example, themicroneedles may be formed of a biodegradable polymer containing adispersion of an active agent for local or systemic delivery. The agentcould be released over time, according to a profile determined by thecomposition and geometry of the microneedles, the concentration of thedrug and other factors. In this way, the drug reservoir is within thematrix of one or more of the microneedles.

In another alternate embodiment, these microneedles may be purposefullysheared off from the substrate after penetrating the biological barrier.In this way, a portion of the microneedles would remain within or on theother side of the biological barrier and a portion of the microneedlesand their substrate would be removed from the biological barrier. In thecase of skin, this could involve inserting an array into the skin,manually or otherwise breaking off the microneedles tips and then removethe base of the microneedles. The portion of the microneedles whichremains in the skin or in or across another biological barrier couldthen release drug over time according to a profile determined by thecomposition and geometry of the microneedles, the concentration of thedrug and other factors. In a preferred embodiment, the microneedles aremade of a biodegradable polymer. The release of drug from thebiodegradable microneedle tips could be controlled by the rate ofpolymer degradation. Microneedle tips could release drugs for local orsystemic effect, but could also release other agents, such as perfume,insect repellent and sun block.

Microneedle shape and content could be designed to control the breakageof microneedles. For example, a notch could be introduced intomicroneedles either at the time of fabrication or as a subsequent step.In this way, microneedles would preferentially break at the site of thenotch. Moreover, the size and shape of the portion of microneedles whichbreak off could be controlled not only for specific drug releasepatterns, but also for specific interactions with cells in the body. Forexample, objects of a few microns in size are known to be taken up bymacrophages. The portions of microneedles that break off could becontrolled to be bigger or smaller than that to prevent uptake bymacrophages or could be that size to promote uptake by macrophages,which could be desirable for delivery of vaccines.

b. Diagnostic Sensing of Body Fluids (Biosensors)

One embodiment of the devices described herein may be used to removematerial from the body across a biological barrier, i.e. for minimallyinvasive diagnostic sensing. For example, fluids can be transported frominterstitial fluid in a tissue into a reservoir in the upper portion ofthe device. The fluid can then be assayed while in the reservoir or thefluid can be removed from the reservoir to be assayed, for diagnostic orother purposes. For example, interstitial fluids can be removed from theepidermis across the stratum corneum to assay for glucose concentration,which should be useful in aiding diabetics in determining their requiredinsulin dose. Other substances or properties that would be desirable todetect include lactate (important for athletes), oxygen, pH, alcohol,tobacco metabolites, and illegal drugs (important for both medicaldiagnosis and law enforcement).

The sensing device can be in or attached to one or more microneedles, orin a housing adapted to the substrate. Sensing information or signalscan be transferred optically (e.g., refractive index) or electrically(e.g., measuring changes in electrical impedance, resistance, current,voltage, or combination thereof). For example, it may be useful tomeasure a change as a function of change in resistance of tissue to anelectrical current or voltage, or a change in response to channelbinding or other criteria (such as an optical change) wherein differentresistances are calibrated to signal that more or less flow of drug isneeded, or that delivery has been completed.

In one embodiment, one or more microneedle devices can be used for (1)withdrawal of interstitial fluid, (2) assay of the fluid, and/or (3)delivery of the appropriate amount of a therapeutic agent based on theresults of the assay, either automatically or with human intervention.For example, a sensor delivery system may be combined to form, forexample, a system which withdraws bodily fluid, measures its glucosecontent, and delivers an appropriate amount of insulin. The sensing ordelivery step also can be performed using conventional techniques, whichwould be integrated into use of the microneedle device. For example, themicroneedle device could be used to withdraw and assay glucose, and aconventional syringe and needle used to administer the insulin, or viceversa.

In an alternate embodiment, microneedles may be purposefully sheared offfrom the substrate after penetrating the biological barrier, asdescribed above. The portion of the microneedles which remain within oron the other side of the biological barrier could contain one or morebiosensors. For example, the sensor could change color as its output.For microneedles sheared off in the skin, this color change could beobserved through the skin by visual inspection or with the aid of anoptical apparatus.

Other than transport of drugs and biological molecules, the microneedlesmay be used to transmit or transfer other materials and energy forms,such as light, electricity, heat, or pressure. The microneedles, forexample, could be used to direct light to specific locations within thebody, in order that the light can directly act on a tissue or on anintermediary, such as light-sensitive molecules in photodynamic therapy.The microneedles can also be used for aerosolization or delivery forexample directly to a mucosal surface in the nasal or buccal regions orto the pulmonary system.

The microneedle devices disclosed herein also should be useful forcontrolling transport across tissues other than skin. For example,microneedles could be inserted into the eye across, for example,conjunctiva, sclera, and/or cornea, to facilitate delivery of drugs intothe eye. Similarly, microneedles inserted into the eye could facilitatetransport of fluid out of the eye, which may be of benefit for treatmentof glaucoma. Microneedles may also be inserted into the buccal (oral),nasal, vaginal, or other accessible mucosa to facilitate transport into,out of, or across those tissues. For example, a drug may be deliveredacross the buccal mucosa for local treatment in the mouth or forsystemic uptake and delivery. As another example, microneedle devicesmay be used internally within the body on, for example, the lining ofthe gastrointestinal tract to facilitate uptake of orally-ingested drugsor the lining of blood vessels to facilitate penetration of drugs intothe vessel wall. For example, cardiovascular applications include usingmicroneedle devices to facilitate vessel distension or immobilization,similarly to a stent, wherein the microneedles/substrate can function asa “staple-like” device to penetrate into different tissue segments andhold their relative positions for a period of time to permit tissueregeneration. This application would be particularly useful withbiodegradable devices. These uses may involve invasive procedures tointroduce the microneedle devices into the body or could involveswallowing, inhaling, injecting or otherwise introducing the devices ina non-invasive or minimally-invasive manner.

The present invention will be further understood with reference to thefollowing non-limiting examples.

EXAMPLE 1

Fabrication of Solid Silicon Microneedles

A chromium masking material was deposited onto silicon wafers andpatterned into dots having a diameter approximately equal to the base ofthe desired microneedles. The wafers were then loaded into a reactiveion etcher and subjected to a carefully controlled plasma based onfluorine/oxygen chemistries to etch very deep, high aspect ratio valleysinto the silicon. Those regions protected by the metal mask remain andform the microneedles.

<100>-oriented, prime grade, 450-550 μm thick, 10-15 Ω-cm silicon wafers(Nova Electronic Materials Inc., Richardson, Tex.) were used as thestarting material. The wafers were cleaned in a solution of 5 parts byvolume deionized water, 1 part 30% hydrogen peroxide, and I part 30%ammonium hydroxide (J. T. Baker, Phillipsburg, N.J.) at approximately80° C. for 15 minutes, and then dried in an oven (Blue M Electric,Watertown, Wis.) at 150° C. for 10 minutes. Approximately 1000 Å ofchromium (Mat-Vac Technology, Flagler Beach, Fla.) was deposited ontothe wafers using a DC-sputterer (601 Sputtering System, CVC Products,Rochester, N.Y.). The chromium layer was patterned into 20 by 20 arraysof 80 μm diameter dots with 150 μm center-to-center spacing using thelithographic process described below.

A layer of photosensitive material (1827 photoresist, Shipley,Marlborough, Mass.) was deposited onto the chromium layer covering thesilicon wafers. A standard lithographic mask (Telic, Santa Monica,Calif.) bearing the appropriate dot array pattern was positioned on topof the photoresist layer. The wafer and photoreist were then exposed toultraviolet (UV) light through the mask by means of an optical maskaligner (Hybralign Series 500, Optical Associates, Inc., Milpitas,Calif.). The exposed photoresist was removed by soaking the wafers in aliquid developer (354 developer, Shipley, Marlborough, Mass.) leavingthe desired dot array of photoresist on the chromium layer.Subsequently, the wafers were dipped into a chromium etchant (CR-75;Cyanteck Fremont, Calif.), which etched the chromium that had beenexposed during the photolithography step, leaving dot arrays of chromium(covered with photoresist) on the surface of the silicon wafer. Thephotoresist still present on the chromium dots formed the masks neededfor fabrication of the microneedles, described below.

The microneedles were fabricated using a reactive ion etching techniquesbased on the Black Silicon Method developed at the University of Twente.The patterned wafers were etched in a reactive ion etcher (700 serieswafer/batch Plasma Processing System, Plasma Therm, St. Petersburg,Fla.) with means for ensuring good thermal contact between the wafersand the underlying platen (Apiezon N, K. J. Lesker, Clairton, Pa.). Thewafers were etched using the following gases and conditions: SF₆ (20standard cubic centimeters per minute) and O₂ (15 standard cubiccentimeters per minute) at a pressure of 150 mTorr and a power of 150 Wfor a run time of approximately 250 minutes. These conditions causedboth deep vertical etching and slight lateral underetching. Bycontrolling the ratio of flow rates of the SF₆ and O₂ gases used to formthe plasma, the aspect ratio of the microneedles could be adjusted. Theregions protected by the chromium masks remained and formed themicroneedles. Etching was allowed to proceed until the masks fell offdue to underetching, resulting in an array of sharp silicon spikes.

EXAMPLE 2

Transdermal Transport Using Solid Microneedles

To determine if microfabricated microneedles could be used to enhancetransdermal drug delivery, arrays of microneedles were made using a deepplasma etching technique. Their ability to penetrate human skin withoutbreaking was tested and the resulting changes in transdermal transportwere measured.

Arrays of microneedles were fabricated having extremely sharp tips(radius of curvature less than 1 μm), and are approximately 150 μm long.Because the skin surface is not flat due to dermatoglyphics and hair,the full length of these microneedles will not penetrate the skin. Allexperiments were performed at room temperature (23±2° C).

The ability of the microneedles to pierce skin without breaking was thentested. Insertion of the arrays into skin required only gentle pushing.Inspection by light and electron microscopy showed that more than 95% ofmicroneedles within an array pierced across the stratum corneum of theepidermis samples. Moreover, essentially all of the microneedles thatpenetrated the epidermis remained intact. On those very few which broke,only the top 5-10 μm was damaged. Microneedle arrays could also beremoved without difficulty or additional damage, as well as re-insertedinto skin multiple times.

To quantitatively assess the ability of microneedles to increasetransdermal transport, calcein permeability of human epidermis with andwithout inserted microneedle arrays was measured. Calcein crosses skinvery poorly under normal circumstances and therefore represents anespecially difficult compound to deliver. As expected, passivepermeability of calcein across unaltered skin was very low, indicatingthat the epidermis samples were intact.

Insertion of microneedles into skin was capable of dramaticallyincreasing permeability to calcein. When microneedles were inserted andleft embedded in the skin, calcein permeability was increased by morethan 1000-fold. Insertion of microneedles for 10 s, followed by theirremoval, yielded an almost 10,000-fold increase. Finally, insertion of amicroneedle array for 1 h, followed by its removal, increased skinpermeability by about 25,000-fold. Permeabilities for skin withmicroneedles inserted and then removed are higher than for skin withmicroneedles remaining embedded probably because the microneedlesthemselves or the silicon plate supporting the array may block access tothe microscopic holes created in the skin. Light microscopy showed thatthe holes which remained in the skin after microneedles were removedwere approximately 1 μm in size.

To confirm in vitro experiments which showed that skin permeability canbe significantly increased by microneedles, studies were conducted withhuman volunteers. They indicated that microneedles could be easilyinserted into the skin of the forearm or hand. Moreover, insertion ofmicroneedle arrays was never reported to be painful, but sometimeselicited a mild “wearing” sensation described as a weak pressure or thefeeling of a piece of tape affixed to the skin. Although transportexperiments were not performed in vivo, skin electrical resistance wasmeasured before and after microneedle insertion. Microneedles caused a50-fold drop in skin resistance, a drop similar to that caused by theinsertion of a 30-gauge “macroneedle.” Inspection of the siteimmediately after microneedle insertion showed no holes visible by lightmicroscopy. No erythema, edema or other reaction to microneedles wasobserved over the hours and days which followed. This indicates thatmicroneedle arrays can permeabilize skin in human subjects in anon-painful and safe manner.

EXAMPLE 3

Fabrication of Silicon Microtubes

Three-dimensional arrays of microtubes were fabricated from silicon,using deep reactive ion etching combined with a modified black siliconprocess in a conventional reactive ion etcher. The fabrication processis illustrated in FIGS. 5a-d. First, arrays of 40 μm diameter circularholes 32 were patterned through photoresist 34 into a 1 μm thick SiO₂layer 36 on a two inch silicon wafer 38 (FIG. 5a). The wafer 38 was thenetched using deep reactive ion etching (DRIE) (Laermer, et al., “BoschDeep Silicon Etching: Improving Uniformity and Etch Rate for AdvancedMEMS Applications,” Micro Electro Mechaiical Systems, Orlando, Fla., USA(Jan. 17-21, 1999)). in an inductively coupled plasma (ICP) reactor toetch deep vertical holes 40. The deep silicon etch was stopped after theholes 40 are approximately 200 μm deep into the silicon substrate 38(FIG. 5b) and the photoresist 34 was removed. A second photolithographystep patterned the remaining SiO₂ layer 36 into circles concentric tothe holes, thus leaving ring shaped oxide masks 34 surrounding the holes(FIG. 5c). The photoresist 34 was then removed and the wafer 38 wasagain deep silicon etched, while simultaneously the holes 40 were etchedcompletely through the wafer 38 (inside the SiO₂ ring) and the siliconwas etched around the Sio₂ ring 38 leaving a cylinder 42 (FIG. 5d). Theresulting tubes were 150 μm in height, with an outer diameter of 80 μm,an inner diameter of 40 μm, and a tube center-to-center spacing of 300μm.

EXAMPLE 4

Micromold Fabrication of Metal Microtubes

Hollow metal microtubes were prepared without dry silicon etching, usinga thick, photo-defined mold of epoxy. The sequences are illustrated inFIGS. 6a-e. First, a thick layer of SU-8 epoxy 44 was spin cast onto asilicon or glass substrate 46 that had been coated with 30 nm oftitanium 48, the sacrificial layer. Arrays of cylindrical holes 49 werethen photolithographically defined through an epoxy layer 44, typically150 μm thick (FIG. 6a). The sacrificial layer then was partially removedusing a wet etching solution containing hydrofluoric acid and water atthe bottom of the cylindrical holes in the SU-8 photoresist 46 (FIG.6b). A seed layer of Ti/Cu/Ti (30 nm/200 nm/30 nm) 39 was thenconformally DC sputter-deposited onto the upper surface of the epoxymold and onto the sidewalls of the cylindrical holes 49 (FIG. 6c). Asshown in FIG. 6c, the seed layer 39 was electrically isolated from thesubstrate. Subsequently, NiFe was electroplated onto the seed layer 39(FIG. 6d), the epoxy 44 was removed from the substrate, and thesurrounding epoxy 44 was removed (FIG. 6e). The resulting microtubes are200 μm in height with an outer diameter of 80 lm, an inner diameter of60 μm, and a tube center-to-center spacing of 150 μm. The holes in theinterior of the microtubes protrude through the base metal supportingthe tubes.

EXAMPLE 5

Micromold Fabrication of Tapered Microneedles

A micromold having tapered walls was fabricated by molding a preexisting3-D array of microneedles, i.e. the mold-insert, and subsequentlyremoving the mold insert. The micromold was then surface plated in amanner similar to that for the microtubes described in Example 4. Thefabrication sequence is illustrated in FIGS. 7a-7 d.

First, an array of solid silicon microneedles 50 were prepared asdescribed in Henry, et al., “Micromachined Needles for the TransdermalDelivery of Drugs,” Micro Electro Mechanical Systems, Heidelberg,Germany, Jan. 26-29, pp. 494-498 (1998). Then, a layer of epoxy 52(SU-8) was spin cast onto the microneedle array to completely blanketthe array (FIG. 7a). The epoxy 52 settled during pre-bake to create aplanar surface above the tips of the microneedles 50. The epoxy 52 wasthen fully pre-baked, photolithographically cross-linked, andpost-baked.

Then, the upper surface of the epoxy 52 was etched away using an O₂/CHF₃plasma until approximately 1 to 2 μm of the needle tips 54 were exposed,protruding from the epoxy 52 (FIG. 7b). The silicon was then selectivelyremoved by using a SF₆ plasma (FIG. 7c). The remaining epoxy mold 52provided a negative of the microneedles with a small diameter hole wherethe tip of the silicon needle protruded. After the removal of thesilicon, a seed layer of Ti—Cu—Ti 54 was conformally sputter-depositedonto the top and sidewalls of the epoxy micromold 52. Following the sameprocess sequence as described in Example 4, NiFe was then electroplatedonto the seed layer 54 (FIG. 7c). Finally, the epoxy was removed usingan O₂/CHF₃ plasma, leaving a 20×20 array of NiFe hollow metalmicroneedles 54 (FIG. 7d). The microneedles 54 were 150 μm in heightwith a base diameter of 80 μm, a tip diameter of 10 μm, and a needle toneedle spacing of 150 μm.

EXAMPLE 6

Micromold Fabrication of Tapered Microneedles Using Laser-Formed Molds

A micromold having tapered walls was fabricated by use of laser ablationtechniques, as shown in FIGS. 8a-d. A laser-ablatable polymer sheet 60such as KAPTON™ polymide approximately 150 microns in thickness wasoptionally laminated to a thin (10-30 micron) metal sheet 62 such astitanium (FIG. 8a). A tapered bole 64 was formed in the metal/polymerlaminate 60/62 using a laser technique such as excimer laser ablation(FIG. 8b). The entry hole of the laser spot was on the metal side 62,and a through hole was made through both the metal sheet and the polymerfilm. The through hole 64 was tapered in combination with eitherdefocusing or appropriate substrate motion to create a taper such thatthe wide end of the hole 64 (typically 40-50 microns) was on the metalside 62 and the narrow end of the hole 64 (typically 10-20 microns) wason the polymer 60 side. A thin layer of metal 66, e.g. titanium, ofthickness 0.1 micron was then deposited, e.g., using asputter-deposition technique, in such a way that the metal 66 depositedon the metal film side and coated the polymer sidewalls, but did notcoat the polymer 60 side of the laminate (FIG. 8c). Electrodeposition ofmetal 68, e.g., gold, to a thickness of 1-5 microns was then performedon the titanium-coated metal surface 66, and polymer sidewalls curvedsection of 60 next to 64. Finally, the polymer 60 was removed, usinge.g. an oxygen plasma, to form the completed microneedles (FIG. 8d).

Alternate polymer removal methods, such as thermal, solvent, aqueous, orphodegradation followed by solvent or aqueous removal, are also possibleif the polymer material is chosen appropriately (e.g., a photoresistresin).

EXAMPLE 7

Formation of Microneedles by Embossing

Formation of a microneedle by embossing is shown in FIGS. 9a-9 f. Apolymeric layer 70 (FIG. 9a) is embossed by a solid microneedle ormicroneedle array 72 (FIG. 9b). The array 72 is removed (FIG. 9c), andthe layer 70 is etched from the non-embossed side 74 until the embossedcavity 76 is exposed (FIG. 9d). A metallic layer 78 is then deposited onthe embossed side and the sidewalls, but not on the non-embossed side 74(FIG. 9e). This layer 78 is optionally thickened by electrodeposition ofan additional metal layer 80 on top of it (FIG. 9e). The polymer layer70 is then removed to form the microneedles 78/80 (FIG. 9f).

EXAMPLE 8

Transdermal Application of Hollow Microneedles

The bore of hollow microneedles must provide fluid flow with minimalclogging in order to be suitable to transport material, such as intransdermal drug delivery. Therefore, microneedies and microtubes wereevaluated to determine their suitability for these functions.

Hollow metal and silicon microneedles, produced as described in Examples3-5, were inserted through human skin epidermis with no apparentclogging of the needle bores. Scanning electron microscopy of a hollowmetal (NiFe) microneedle penetrating up through the underside of humanepidermis showed the microneedle remains intact, with the tip free ofdebris. Similarly, silicon microneedles, metal microneedles, and metalmicrotubes were successfully inserted through human skin. Also, thehollow microneedles were shown to permit the flow of water through theirbores.

EXAMPLE 9

Transport of Drugs through Microneedles Inserted into Skin

Studies were performed with solid and hollow microneedles to demonstratetransport of molecules and fluids. As shown in Table 1, transport of anumber of different compounds across skin is possible usingmicroneedles. These studies were performed using either solid siliconmicroneedles or using hollow silicon microneedles made by methodsdescribed in this patent. Transport was measured across human cadaverepidermis in vitro using Franz diffusion chambers at 37° C. usingmethods described in S. Henry, D. McAllister, M. G. Allen and M. R.Prausnitz. Microfabricated microneedles: A novel method to increasetransdermal drug delivery. J. Pharm. Sci. 87, 922-925 (1998).

The transdermal delivery of calcein, insulin, bovine serum albumin andnanoparticles was measured. Delivery refers to the ability to transportthese compounds from the stratum corneum side of the epidermis to theviable epidermis side. This is the direction of transport associatedwith delivering drugs into the body. Removal of calcein was alsomeasured. Removal refers to the ability to transport calcein from theviable epidermis side of the epidermis to the stratum corneum side. Thisis the direction of transport associated with removing from the bodycompounds found in the body, such as glucose.

In all cases shown in Table 1, transport of these compounds across skinoccurred at levels below our detection limit when no needles wereinserted into the skin. Intact skin provides an excellent barrier totransport of these compounds. In all cases examined, when solidmicroneedles were inserted into the skin and left in place, large skinpermeabilities were measured, indicating that the microneedles hadcreated pathways for transport across the skin. Furthermore, in allcases, when solid microneedles were inserted into the skin and thenremoved, even greater skin permeabilities resulted. Finally, when hollowmicroneedles were inserted into the skin and left in place, stillgreater skin permeabilities resulted for those compounds tested. Thesestudies show that microneedles can dramatically increase skinpermeability and can thereby increase transport of a number of differentcompounds across the skin. It also shows that when solid microneedlesare used, a preferred embodiment involves inserting and then removingmicroneedles, rather than leaving them in place. It also shows thatusing hollow microneedles are a preferred embodiment over the use ofsolid microneedles.

In Table 2, the flow rate of water through hollow silicon microneedlesis shown as a function of applied pressure. These data demonstrate thatsignificant flow rates of water through microneedles can be achieved atmodest pressures.

TABLE 1 Transport of Drugs through Microneedles inserted into Skin.Solid needles No Solid needles inserted and Hollow needles Compoundneedles inserted removed inserted Calcein delivery ** 4 × 10⁻³ 1 × 10⁻²1 × 10⁻¹ Calcein removal ** 2 × 10⁻³ 1 × 10⁻² n.a. Insulin delivery ** 1× 10⁻⁴ 1 × 10⁻² n.a. Bovine serum ** 9 × 10⁻⁴ 8 × 10⁻³ 9 × 10⁻² albumindelivery Nanoparticle ** n.a. 3 × 10⁻⁵ n.a. delivery **means that thetransport was below the detection limit. n.a. means that the data arenot available. Nanoparticles were made of latex with a diameter ofapproximately 100 nm.

TABLE 2 Flow rate of water through hollow silicon microneedles as afunction of applied pressure Pressure (psi) Flow rate (ml/min) 1.0 161.5 24 2.0 31 2.5 38 3.0 45

Publications cited herein and the material for which they are cited arespecifically incorporated by reference.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

We claim:
 1. A method for making a microneedle device having a substrateto which a plurality of microneedles are attached or integrally formed,the method comprising (i) forming a micromold having sidewalls whichdefine a surface of each of the microneedles; and (ii) using themicromold to form the microneedles and the substrate, wherein themicroneedles extend at an angle from the substrate.
 2. The method ofclaim 1 wherein the micromold is formed by a process which includesproviding a mold substrate and then forming a plurality of holes in themold substrate, thereby forming the micromold wherein the inner surfacesof the holes define the sidewalls which correspond to outer surfaces ofeach of the microneedles.
 3. The method of claim 2 wherein the holes arephotolithographically defined.
 4. The method of claim 2 wherein theholes are defined using a laser technique.
 5. The method of claim 4wherein the laser technique is laser ablation.
 6. The method of claim 1comprising, in step (ii), applying a metal, or other material havingdifferent properties than the material forming the micromold, to thesidewalls to form hollow microneedles, and then removing the micromoldfrom the microneedles.
 7. The method of claim 6 wherein the metal orother material is applied by a sputter-deposition technique or aelectrodeposition technique.
 8. The method of claim 6 wherein themicromold is removed using a process selected from the group consistingof oxygen plasma, thermal, solvent, aqueous, and photodegradationremoval techniques.
 9. The method of claim 6 wherein the metal or othermaterial is applied by electroless plating.
 10. The method of claim 1comprising, in step (ii), filling the micromold with a fluid materialthat is hardened in the micromold to form the microneedles.
 11. Themethod of claim 10 which utilizes injection molding or reactioninjection molding.
 12. The method of claim 1 wherein, in step (i), themicromold is fabricated by forming a mold from a mold-insert.
 13. Themethod of claim 12 wherein the mold insert is an array of microneedles.14. The method of claim 13 for forming hollow microneedles, comprisingthe steps of (a) layering a removable material onto the array to coverthe microneedles of the mold-insert, (b) removing a part of the layer ofremovable material to expose the tips of the microneedles of themold-insert, and (c) removing the mold-insert to yield a micromold. 15.The method of claim 14 further comprising (d) applying a metal, or othermaterial having properties distinct from the material forming themicromold, onto the micromold to form the microneedles, and (e) removingthe micromold from the microneedles.
 16. The method of claim 14 whereinthe micromold is removed using a process selected from the groupconsisting of oxygen plasma, thermal, solvent, aqueous, andphotodegradation removal techniques.
 17. The method of claim 1 whereinthe micromold is shaped by embossing.
 18. The method of 1 wherein themicromold is shaped using a laser to selectively remove material. 19.The method of claim 18 wherein the material is a polymer/metallaminante.
 20. The method of claim 1 wherein the angle is about 90°. 21.The method of claim 2 wherein the microneedle is formed of a metalselected from the group consisting of nickel, iron, gold, titanium,copper, platinum, palladium, stainless steels, and alloys thereof. 22.The method of claim 1 wherein the micromold is formed of a polymericmaterial.
 23. The method of claim 1 wherein the microneedles comprise apolymer.
 24. The method of claim 1 wherein the micromold formed in step(i) is in the form of an array of mold microneedles, wherein the outersurfaces of each of the mold microneedles correspond to the sidewallswhich define the inner surface of hollow microneedles; and wherein, instep (ii), the array is coated with a material forming the microneedlesand the substrate of the device, and then the micromold is removed.